Universiti Tunku Abdul Rahman (Kampar Campus) Faculty of Science, Engineering, and Technology Bachelor of Science (Hons) Biotechnology

Year 2 Semester 1 UESB 2142 Laboratory 2A (III) Microbiology Lecturer: Dr. Teh Yok Lan Student’s Name: Cheah Hong Leong Student’s ID: 08AIB03788 Group Member: Samuel Tan Sha Loong Experiment No. 5 Title: Growth Curve Date: August 17, 2009

Title: Growth Curve Objectives: – To obtain the graphs of the microbial growth of different bacteria through optical density measurement of the media. – To compare the growth curve of the bacteria.

Results:

Table 1: Optical Density of Inoculated Nutrient Broth with Time of Pseudomonas aeruginosa Time (minutes) To, 0 T1, 15 T2, 30 T3, 45 T4, 60 T5, 75 T6, 90 Optical Density, OD600 (A) 0.013 0.015 0.017 0.020 0.022 0.026 0.031

Table 2: Optical Density of Inoculated Nutrient Broth with Time of Escherichia coli Time (minutes) To, 0 T1, 15 T2, 30 T3, 45 T4, 60 T5, 75 T6, 90 Optical Density, OD600 (A) 0.023 0.020 0.074 0.085 0.226 0.262 0.223

Table 3: Optical Density of Inoculated Nutrient Broth with Time of Staphylococcus aureus Time (minutes) To, 0 T1, 15 T2, 30 T3, 45 T4, 60 T5, 75 T6, 90 Optical Density, OD600 (A) 0.022 0.028 0.034 0.040 0.062 0.091 0.143

Note: Data in Table 1, 2, and 3 were plotted in “Semi-log Graph: Optical Density of Inoculated Nutrient Broth versus Time” in Attachment. Calculation of Generation Time:

Pseudomonas aeruginosa, Generation time = 45 minutes Escherichia coli, Generation time = T0.100 – T0.050 = (45 – 24) minutes = 21 minutes Staphylococcus aureus, Generation time = T0.100 – T0.050 = (87 – 48) minutes = 39 minutes

Discussion: Based on the semi-log graph and the calculation, the generation time of Escherichia coli was the shortest (21 minutes), followed by Staphylococcus aureus (39 minutes) and Pseudomonas aeruginosa (45 minutes). With the shortest generation time, Escherichia coli was expected to have the greatest growth rate under similar condition among the three microorganisms, for every 21 minutes, the cell numbers of Escherichia coli doubled. Theoretically, the photospectrometry measurement assumes that the bacterial cells were distributed in the nutrient broth evenly; the optical density obtained can be proportional to the population of the bacteria in broth only when the assumption occurred. However, all the three bacteria have the ability to form biofilm on the flask surface, which can cause errors in the photospectrometry measurement regarding the total cell mass of the microorganisms in the nutrient broth.

Stirring of the nutrient broth during the incubation was solving the problem, but during the incubation in the laboratory practical, non simultaneous timing of the students from different subgroups made the incubation machine had to be turned on and off repeatedly. That might lead to the uneven distribution of the bacteria in the broth. Moreover, sometimes the incubation machine was also did not turned on by the students after placing the flask into it, leaving the broth unstirred for several minutes. This might be able to explain the unusual results shown by the Escherichia coli in which the optical density was unexpectedly dropped at time 15 minute and 90 minute. Conclusion: The generation time was shortest for Escherichia coli, followed by Staphylococcus aureus and Pseudomonas aeruginosa.

References: Cappuccino, J. G. & Sherman, N. (2008). Microbiology. A Laboratory Manual, 8th ed., San Francisco, CA: Pearson Benjamin Cummings. Escherichia coli. (2009, August 30). In Wikipedia, the free encyclopedia. Retrieved August 31, 2009, from http://en.wikipedia.org/wiki/Escherichia_coli Madigan, M. T., Martinko, J. M., Dunlap, P. V., & Clark, D. P. (2009). Brock Biology of Microorganisms, 12th ed., San Francisco, CA: Pearson Benjamin Cummings. Pseudomonas aeruginosa. (2009, August 28). In Wikipedia, the free encyclopedia. Retrieved August 31, 2009, from http://en.wikipedia.org/wiki/Pseudomonas_aeruginosa Staphylococcus aureus. (2009, August 26). In Wikipedia, the free encyclopedia. Retrieved August 31, 2009, from http://en.wikipedia.org/wiki/Staphylococcus_aureus

Attachment:

Semi-log Graph: Optical Density of Inoculated Nutrient Broth versus Time

Sign up to vote on this title
UsefulNot useful